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文献和实验Transfecting Suspension Cells(悬浮细胞转染)
in 28 ml of FreeStyle™ 293 Expression Medium. Expand cells accordingly, taking into account the cell doubling time. For FreeStyle™ 293-F cells, this equates to passing cells at ~6–7 × 10 5 cells/ml. 2.On the day of transfection
Mammalian Membrane Protein Expression in Baculovirus-Infected Insect Cells
A system of expression of mammalian membrane proteins in baculovirus-infected insect cells is described, allowing analytical or preparative production in the milligram range of such type of proteins. This is illustrated by the setup
with the better the chances of a successful pulldown. A total of 4.0×106 293 HEK cells (~70% confluent) are plated and 24 hours later transfected with 15µg of a RNA Polymerase II expressing flag-tagged construct plasmid. We generally use Lipofectamine 2000™
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